[Suitability of human settlement environment in Buyei traditional villages in rocky desertification area of Guizhou, China.] / è´µå·žçŸ³æ¼ åŒ–åŒºå¸ƒä¾æ—ä¼ ç»Ÿæ‘è½äººå± çŽ¯å¢ƒé€‚å®œåº¦.

To investigate the traditional village living environment adaptability to desertification and topography, the suitability evaluation index system and weight of traditional Buyei traditional villages in the rocky desertification area human settlement were quantified using Delphi method and comprehensive weighting method. We calculated human settlements environment suitability value and threshold to comprehensively analyze the human settlements environment suitability. The results showed that 10% of the traditional Buyei traditional villages were located in the rocky desertification areas with high intensity and extremely high intensity and had the tradition of selecting the best environment. The index system of residential environment suitability was composed of five first-level indices (economy, historic culture, ecology, society, and building environment suitability) and 26 second-level indices. This index system was applicable to villages in karst regions. The comprehensive environmental suitability value (2.81-3.77), the economy value (0.77-1.17), the historic culture value (0.39-0.50), the ecology value (0.83-1.07), the social environment value (0.38-0.53) all decreased with the increasing intensity of rocky desertification, but the suitability value of building environment did not change, which ranged from 0.43 to 0.51. Rocky desertification had profound and synergistic impacts on economy, historic culture, ecology and social environment. The floor level of the human settlement suitability threshold was 2.93. If the threshold was lower than 2.93, it could be considered to move or take measures to improve its value. The suitability value (3.56) of traditional village living environment in mountain slope was higher than that in depression (3.42) and valley (3.16). The human settlement suitability of traditional villages in rocky desertification area was higher than that of ordinary villages, but was lower than that of normal landform, with the differences in economy and ecology being the main reasons. To improve the living environment of traditional villages in rocky desertification areas, we should strengthen the comprehensive control of rocky desertification and policy support, develop ecological economy and tourism, protect historic culture or choose ecological migration. This research could provide theoretical base for the planning and construction of village living environment protection in karst areas.

Betaine-assisted recombinase polymerase assay with enhanced specificity.

Recombinase polymerase amplification (RPA) is a widespread isothermal amplification method and regarded as an excellent candidate to replace polymerase chain reaction. However, the specificity of RPA is not always satisfactory when the sample contains amounts of background DNA. Herein, we report a novel RPA method named betaine-assisted RPA (B-RPA) that uses inexpensive betaine to avoid nonspecific amplification effectively. Result show that nonspecific amplification is prone to occur in RPA if the primers have not been rigorously refined, especially in detecting samples with large amounts of background DNA. This problem has been addressed by adding betaine to the RPA reactions. Our data show that the addition of 0.8 M betaine can significantly increase specificity and efficiency simultaneously. This B-RPA method is also used to detect hepatitis B virus DNA in clinical plasma samples, thereby demonstrating the clinical practicability of B-RPA.

CD45+CD33lowCD11bdim myeloid-derived suppressor cells suppress CD8+ T cell activity via the IL-6/IL-8-arginase I axis in human gastric cancer.

Myeloid-derived suppressor cells (MDSCs) are a prominent component of the pro-tumoral response. The phenotype of and mechanisms used by MDSCs is heterogeneous and requires more precise characterization in gastric cancer (GC) patients. Here, we have identified a novel subset of CD45+CD33lowCD11bdim MDSCs in the peripheral blood of GC patients compared to healthy individuals. CD45+CD33lowCD11bdim MDSCs morphologically resembled neutrophils and expressed high levels of the neutrophil marker CD66b. Circulating CD45+CD33lowCD11bdim MDSCs effectively suppressed CD8+ T cells activity through the inhibition of CD8+ T cell proliferation and interferon-γ (IFN-γ) and granzyme B (GrB) production. The proportion of CD45+CD33lowCD11bdim MDSCs also negatively correlated with the proportion of IFN-γ+CD8+ T cell in the peripheral blood of GC patients. GC patient serum-derived IL-6 and IL-8 activated and induced CD45+CD33lowCD11bdim MDSCs to express arginase I via the PI3K-AKT signaling pathway. This pathway contributed to CD8+ T cell suppression as it was partially rescued by the blockade of the IL-6/IL-8-arginase I axis. Peripheral blood CD45+CD33lowCD11bdim MDSCs, as well as IL-6, IL-8, and arginase I serum levels, positively correlated with GC progression and negatively correlated with overall patient survival. Altogether, our results highlight that a subset of neutrophilic CD45+CD33lowCD11bdim MDSCs is functionally immunosuppressive and activated via the IL-6/IL-8-arginase I axis in GC patients.

Comparison of the myocardial protective effect of sevoflurane versus propofol in patients undergoing heart valve replacement surgery with cardiopulmonary bypass.

BACKGROUND: This study aimed to compare myocardial protective effects of anaesthesia with intravenous infusion of propofol versus inhalation of sevoflurane in patients undergoing heart valve replacement surgery with cardiopulmonary bypass. METHODS: Seventy-six patients undergoing valve replacement with cardiopulmonary bypass were randomly assigned to propofol or sevoflurane anesthesia during the surgery, respectively. For assessing myocardial injury, cardiac troponin I (cTnI) and creatine kinase isozyme (CK-MB) were determined before induction (T0), 0.5 h (T1) and 3 h (T2) after aortic unclamping, and 24 h (T3) and 48 h (T4) after surgery. The concentrations of interleukin (IL)-6 and IL-10 as the systemic inflammatory and anti-inflammatory markers were also measured at above time points. RESULTS: In the sevoflurane group, the plasma concentrations of cTnI and CK-MB from Tl to T4 and the levels of IL-6 and IL-10 from T1 to T2 were lower than those in the propofol group. Moreover, a higher ratio of automatic heart beat recovery and a shorter length of intensive care unit or hospital stay were found in the sevoflurane group comparing with the propofol group. CONCLUSION: Sevoflurane anaesthesia produced more prominent myocardial protection and attenuated inflammatory response than propofol anaesthesia in patients with valve replacement surgery under cardiopulmonary bypass, resulting in shorter ICU and in-hospital stay. RETROSPECTIVE CLINICAL TRIAL REGISTRATION: Identified as ChiCTR-IOR-16009979 at http://www.chictr.org.cn/ .

Substitution of the precursor peptide prevents anti-prM antibody-mediated antibody-dependent enhancement of dengue virus infection.

Antibody-dependent enhancement (ADE) is currently considered as the mechanism underlying the pathogenesis of severe dengue disease. Many studies have shown that precursor (pr) peptide-specific antibodies do not efficiently neutralize infection but potently promote ADE of dengue virus (DENV) infection. To explore the effect of pr peptide substitution on neutralization and ADE of DENV infection, the rabbit anti-prM polyclonal antibodies (pAbs) and anti-JEVpr/DENV-M pAbs were prepared, and the neutralization and ADE of these two pAbs were further compared. Here, we report that both anti-JEVpr/DENV-M and anti-prM pAbs exhibited broad cross-reactivity and only partial neutralization with four DENV serotypes and immature DENV. Rabbit anti-prM pAbs showed a significant enhancement in a broad range of serum dilutions. However, there was no statistically significant difference in the enhancing activity of rabbit anti-JEVpr/DENV-M pAbs at various levels of dilution. These results demonstrate that anti-prM antibody-mediated ADE can be prevented by JEV pr peptide replacement. The present study contribute further to research on the pathogenesis of DENV infection.

Association of Toll-like receptor 2 polymorphisms with gout.

Gout is the most common autoinflammatory arthritis characterized by elevated serum urate and recurrent attacks of intra-articular crystal deposition of monosodium urate (MSU) in tissues. The pathogenesis of gout has not been fully determined, although certain genetic factors are involved in the development of gout. Accumulated data suggested that MSU crystal-induced inflammation is a paradigm of innate immunity. As Toll-like receptors (TLRs) are the underlying mechanisms of the innate immune response, the present study aimed to investigate whether TLR2 polymorphisms are associated with gout. Two single-nucleotide polymorphisms (Arg677Trp and Arg753Gln, rs5743708) in TLR2 were genotyped by polymerase chain reaction-restriction fragment length polymorphism and the -196 to -174 del polymorphism was investigated using the allele-specific polymerase chain reaction in 431 individuals (215 patients with gout and 216 healthy controls). TLR2 Arg677Trp and Arg753Gln genotyping indicated that all the positive samples were of the wild-type genotype. No significant differences in genotype (χ2=1.686, P=0.430) and allele (χ2=1.430, P=0.232) frequencies of the -196 to -174 del polymorphism between the patients with gout and the control groups was observed. Our results suggested that the TLR2 Arg677Trp, Arg753Gln and the -196 to -174 del polymorphisms were not associated with susceptibility to primary gouty arthritis.

[Role of store-operated Ca2+ channels in primary hepatocytes under conditions of calcium overload and ethanol-induced injury].

OBJECTIVE: To investigate the role of store-operated calcium channels (SOCs) in primary hepatocytes under conditions of calcium overload and ethanol-induced injury. METHODS: The in vitro model of chronic ethanol-induced hepatocyte injury was established using primary hepatocytes isolated from Sprague-Dawley rats. Ethanol-induced changes (24, 48 and 72 h; 50, 100, 200, 400 and 800 mmol/L) in expression of the SOCs proteins stromal interaction molecule 1 (STIM1) and calcium release-activated calcium channel protein 1 (Oria1) were detected by qualitative PCR analysis (mRNA) and western blotting (protein). The possible role of these two SOCs proteins in the ethanol-induced extracellular calcium influx and related liver cell injury was determined by treating the cell system with various channel blockers (EGTA, La3+, and 2-APB). Cell viability was determined by MTT assay and cytosolic free calcium ion concentration was determined by flow cytometry. RESULTS: After 24 h of exposure to 0 (untreated) to 800 mM/L ethanol, the cell viability was reduced in a concentration-dependent manner. The 400 mmol/L concentration of ethanol decreased cell viability by 57.34% +/- 2.34%. and was chosen for use in subsequent experiments. Compared with the untreated control cells, the ethanol-treated cells showed significantly up-regulated mRNA and protein expression of both STIM1 and Orai1 at all times examined, suggesting that the ethanol-stimulated expression of STIM1 and Orai1 could persist for at least 72 h. The ethanol treatment induced increase in cytoplasmic calcium levels was significantly (and similarly) reduced by co-treatment with any of the three channel blockers. CONCLUSION: Chronic ethanol exposure can increase the expression of STIM1 and Orai1 in primary liver cells, suggesting that ethanol may increase extracellular calcium influx by up-regulating expression of these SOCs protein molecules, ultimately aggravating liver cell damage.

Expression of CC chemokine ligand 5 in patients with rheumatoid arthritis and its correlation with disease activity and medication.

OBJECTIVE: To determine the levels of CC chemokine ligand 5 (CCL5) in serum and synovial fluid (SF) from patients with rheumatoid arthritis (RA) and their relations with disease activity and medication. METHODS: CCL5 in serum and SF was quantified by enzyme-linked immunosorbent assay (ELISA) in 28 RA patients and 21 osteoarthritis (OA) patients. In RA patients, the correlations of CCL5 levels in serum and SF with disease activity were analyzed. Meanwhile, the serum CCL5 levels among RA patients treated with disease-modifying antirheumatic drugs (DMARDs), Tripterygium Glucosides, and other Chinese herbs without disease-modifying effects were also compared. RESULTS: CCL5 levels in both serum and SF of RA patients were significantly higher than those of OA patients (P < 0.05). Moreover, the level of CCL5 was higher in SF than that in serum of RA patients (P < 0.01). Serum CCL5 level was correlated significantly with the number of swollen joints (r = 0.3329, P < 0.05), erythrocyte sedimentation rate (r = 0.4001, P < 0.05), and C reactive protein (r = 0.3735, P < 0.01). In addition, the level of CCL5 had a trend of lower in patients treated with DMARDs or Tripterygium Glucosides than those treated with other Chinese herbs, although the difference was not significant among those patients due to the small number of patients in each group. CONCLUSIONS: In RA patients, the expression of CCL5 increases and correlates with some clinical and laboratory parameters of RA, which indicate that CCL5 plays an important role in RA and may serve as a useful marker of disease activity. DMARDs and Tripterygium Glucosides might exert their clinical effects through reducing CCL5 production in RA.

[Membrane and cytoplasmic expression of urokinase-type plasminogen activator receptor in synovial tissues of rheumatoid arthritis patients].

OBJECTIVE: To investigate the production of membrane-expressed urokinase-type plasminogen activator receptor (muPAR) and cytoplasmic-expressed uPAR (cuPAR) in synovial tissue from patients with rheumatoid arthritis (RA) and analyze their relationship with the severity of synovial inflammation in RA. METHODS: MuPAR and cuPAR were measured with indirect immunofluorescence (IIF) and immunoperoxidase histochemical analysis of the synovial tissue sections from 18 patients with RA, 10 patients with osteoarthritis (OA) and 4 healthy subjects. The association of uPAR expression with the severity of synovitis in RA was then analyzed. RESULTS: MuPAR positive cells were detected in approximately (66.0+/-9.4)% of the RA synovial cells, distributed predominantly in vascular endothelial cells and fibroblast cells. While cuPAR positive cells were found in about (61.0+/-5.8)% of the RA synovial cells, including subsynovial and interstitial macrophage-like cells, mononuclear leukocytes and fibroblast cells. Both the muPAR and cuPAR expression were much more increased in RA synovial tissue than those in OA synovial tissue. Furthermore, the number of muPAR (r=0.672, P<0.01) and cuPAR (r=0.649, P<0.01) positive cells in synovial tissue was also found to be correlated significantly with the severity of synovial inflammation in RA patients. CONCLUSION: The up-regulated expression of muPAR in synovial vascular endothelial cells suggests an important role of this molecule in angiogenesis in RA and the increased production of cuPAR in synovial inflammatory cells indicates the involvement of cuPAR in the inflammatory process in RA.

[Construction and expression of fusion gene expression plasmid HBV preS2S-rhGM-CSF].

AIM: To construct GM-CSF and preS2 fusion gene expression plasmid to enhance the immunogenicity of hepatitis B DNA vaccine. METHODS: HBV preS2S gene(846 bp) and rhGM-CSF gene(384 bp) were amplified by PCR, respectively. The eukaryotic expression plasmid pcDNA3.1-S2S-rhGM-CSF was constructed by means of T-A clone and directional gene cloning techniques, and then the recombinant plasmid was expressed in HepG2 cells. RESULTS: Restriction enzyme digestion analysis, PCR amplification and/or DNA sequencing proved that the recombinant plasmid was constructed successfully. The transcription of target gene was confirmed by RT-PCR. The fusion protein expressed in HepG2 cells could reacted to the monoclonal antibodies (mAbs) against HBsAg, preS2 and GM-CSF, respectively. CONCLUSION: The successful construction and expression of pcDNA3.1-S2S-rhGM-CSF lay the foundation for further study of HBV DNA vaccine.